Investigating the Antitumor Effects of α-Lactalbumin–Oleic Acid Complexes (HAMLET and BAMLET) in Colorectal Cancer: A Systematic Perspective
Abstract
Objective: Recently, attention has turned toward naturally derived complexes such as HAMLET (Human α-lactalbumin Made Lethal to Tumor Cells) and BAMLET (Bovine α-lactalbumin Made Lethal to Tumor Cells) due to their selective cytotoxicity and promising mechanisms of action. In this study, we aimed to systematically review the effect of HAMLET and BAMLET on colorectal cancer (CRC).
Methods: PubMed, Scopus, Web of Science, and Embase were searched for studies published between 2020–2025, with one 2006 study included for its unique mechanistic insights. The search combined specific keywords and MeSH terms related to HAMLET, BAMLET, colorectal cancer, antitumor activity, and signaling pathways, using Boolean operators. After removing duplicates and applying predefined inclusion and exclusion criteria, six eligible original studies (in vitro, in vivo, and xenograft models) were included.
Results: Both HAMLET and BAMLET demonstrated cytotoxic effects against CRC cells via diverse mechanisms such as autophagy inhibition, lysosomal membrane permeabilization, mitochondrial dysfunction, and modulation of β-catenin and CK1α signaling. BAMLET notably exhibited synergistic effects with 5-fluorouracil and contributed to reduced tumor growth in murine models. HAMLET’s efficacy varied depending on KRAS/BRAF mutation status and mitochondrial resilience. Importantly, no significant toxicity was observed in healthy non-cancerous cells across the studies.
Conclusions: These findings suggest that HAMLET and BAMLET represent viable adjuncts to standard CRC therapies, offering tumor-specific mechanisms with minimal side effects. Further exploration of their molecular interactions and clinical potential could enhance combination strategies and help overcome therapeutic resistance in CRC treatment.
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5. Jafari A, Hosseini FA, Jalali, FS. A systematic review of the economic burden of colorectal cancer. Health Sci. Rep. 2024; 7(8), e70002. https://doi.org/10.1002/hsr2.70002.
6. Asma ST, Acaroz U, Imre K, Morar A, Shah SRA, Hussain SZ, et al. Natural Products/Bioactive Compounds as a Source of Anticancer Drugs. Cancers. 2022; 14(24):6203. https://doi.org/10.3390/cancers14246203.
7. Håkansson A, Zhivotovsky B, Orrenius S, Sabharwal H, Svanborg C. Apoptosis induced by a human milk protein. Proc Natl Acad Sci U S A. 1995; 92(17):8064-8. https://doi.org/10.1073/pnas.92.17.8064.
8. Wang M, Liu X, Chen T, Cheng X, Xiao H, Meng X, Jiang Y. Inhibition and potential treatment of colorectal cancer by natural compounds via various signaling pathways. Front Oncol. 2022; 12:956793.9. https://doi.org/10.3389/fonc.2022.956793.
9. Islam MR, Akash S, Rahman MM, Nowrin FT, Akter T, Shohag S, et al. Colon cancer and colorectal cancer: Prevention and treatment by potential natural products. Chem Biol Interact. 2022; 368:110170. https://doi.org/10.1016/j.cbi.2022.110170.
10. Zacatecas Z, Campus, II. Perspective of the Use of Hamlet as Palliative in Patients Undergoing Chemotherapy. Acta Sci. Med. Sci. 2024; 8(7). https://doi.org/10.31080/ASMS.2024.08.1842
11. Ho JC, Nadeem A, Svanborg C. HAMLET–A protein-lipid complex with broad tumoricidal activity. Biochem Biophys Res Commun. 2017; 482(3), 454-458. https://doi.org/10.1016/j.bbrc.2016.10.092.
12. Žilinskas J, Stukas D, Jasukaitienė A, Šapauskienė J, Banienė R, Trumbeckaitė S, et al. HAMLET effect on cell death and mitochondrial respiration in colorectal cancer cell lines with KRAS/BRAF mutations. J. Cancer Res. Clin. Oncol. 2023; 149(11):8619-8630. https://doi.org/10.1007/s00432-023-04777-0
13. Žilinskas J. HAMLET Complex and FOLFOX Chemotherapy Impact on Colorectal Cancer Cells and Tissues with Different KRAS/BRAF Mutations. Doctoral dissertation, Lithuanian University of Health Sciences (Lithuania)). 2024.
14. Babazadeh M, Zamani M, Mehrbod P, Mokarram P. The Comparative Role of BAMLET and 5-Fluorouracil in Colorectal Cancer Cells by Targeting WNT/β-Catenin Pathway.Int J Cancer Manag. 2022; 15(9):e123140. https://doi.org/10.5812/ijcm-123140.
15. Zhao H, Ming T, Tang S, Ren S, Yang H, Liu M, et al. Wnt signaling in colorectal cancer: pathogenic role and therapeutic target. Mol Cancer. 2022; 21(1), 144. https://doi.org/10.1186/s12943-022-01616-7.
16. Behrouj H, Mokarram P. BAMLET (Bovine α-lactalbumin made lethal to tumor cells) inhibits autophagy flux and induces apoptosis via down-regulation of protein kinase CK1α and attenuation of the AKT/p-ß-catenin (S552) pathway in RAS-mutated human colorectal HCT 116 cells. Iran J Basic Med Sci. 2023; 26(10):1212-1219. https://doi.org/10.22038/IJBMS.2023.69343.15114.
17. Puthia M, Storm P, Nadeem A, Hsiung S, Svanborg C. Prevention and treatment of colon cancer by peroral administration of HAMLET (human α-lactalbumin made lethal to tumour cells). Gut. 2014; 63(1):131-142. https://doi.org/10.1136/gutjnl-2012-303715.
18. Payandeh Z, Khalili S, Somi MH, Mard-Soltani M, Baghbanzadeh A, Hajiasgharzadeh K, et al. PD-1/PD-L1-dependent immune response in colorectal cancer. J. Cell. Physiol. 2020; 235(7-8), 5461–5475. https://doi.org/10.1002/jcp.29494.
19. Dosset M, Vargas TR, Lagrange A, Boidot R, Végran F, Roussey A, et al. PD-1/PD-L1 pathway: an adaptive immune resistance mechanism to immunogenic chemotherapy in colorectal cancer. Oncoimmunol, 2018; 7(6), e1433981. https://doi.org/10.1080/2162402X.2018.1433981.
20. Tran HT, Wan MLY, Ambite I, Cavalera M, Grossi M, Háček J, et al. BAMLET administration via drinking water inhibits intestinal tumor development and promotes long-term health. Sci Rep. 2024; 14, 3838. https://doi.org/10.1038/s41598-024-54040-w.
21. Jiang H, Pang J, Li T, Akofala A, Zhou X, Yi C, et al. PD-1 regulates the anti-tumor immune function of macrophages through JAK2-STAT3 signaling pathway in colorectal cancer tumor microenvironment. J Transl Med. 2025; 23(1):502. https://doi.org/10.1186/s12967-025-06469-4
22. & Svanborg, C. (2008). Apoptosis and tumor cell death in response to HAMLET (human α-lactalbumin made lethal to tumor cells). Bioactive components of milk, 217-240.
23. Hallgren O, Aits S, Brest P, Gustafsson L, Mossberg AK, Wullt B, et al. Apoptosis and Tumor Cell Death in Response to HAMLET (Human α-Lactalbumin Made Lethal to Tumor Cells). In: Bösze, Z. (eds) Bioactive Components of Milk. Advances in Experimental Medicine and Biology, vol 606. Springer, New York, NY. 2008; https://doi.org/10.1007/978-0-387-74087-4_8.
24. Hallgren O, Gustafsson L, Irjala H, Selivanova G, Orrenius S, Svanborg C. HAMLET triggers apoptosis but tumor cell death is independent of caspases, Bcl-2 and p53. Apoptosis. 2006; 11, 221–233. https://doi.org/10.1007/s10495-006-3607.
25. Žilinskas J, Stukas D, Jasukaitienė A, Žievytė I, Balion Z, Šapauskienė J, et al. Assessing the Therapeutic Impacts of HAMLET and FOLFOX on BRAF-Mutated Colorectal Cancer: A Study of Cancer Cell Survival and Mitochondrial Dynamics in Vitro and Ex Vivo. Medicina. 2024; 60(1):142. https://doi.org/10.3390/medicina60010142.
26. Babazadeh M, Zamani M, Mehrbod P, Mokarram P. Stemness targeting of colorectal cell lines mediated by BAMLET and 5-Flourouracil. Biochem Biophys Res Commun. 2023; 664:136-141. https://doi.org/10.1016/j.bbrc.2023.04.010.
27. Rammer P, Groth-Pedersen L, Kirkegaard T, Daugaard M, Rytter A, Szyniarowski P, et al. BAMLET activates a lysosomal cell death program in cancer cells. Mol. Cancer Ther. 2010; 9(1), 24-32. https://doi.org/10.1158/1535-7163.mct-09-0559.
28. Wu X, Que H, Li Q, Wei X. Wnt/β-catenin mediated signaling pathways in cancer: recent advances, and applications in cancer therapy. Mol. Cancer. 2025; 24(1):171. Published 2025 Jun 10. http://doi:10.1186/s12943-025-02363-1.
29. Yang Z, Manshour N, He X, Epp M, Unruh J, Mao X, et al. Overcoming Immune Escape Via Targeting Akt-Wnt/β-Catenin By Artificial-Intelligence-Powered Peptide-Producing mRNAs. 2024; Blood, 144, 1062. https://doi.org/10.1182/blood-2024-200368.
30. Hoque M, Dave S, Gupta P, Saleemuddin M. Oleic acid may be the key contributor in the BAMLET-induced erythrocyte hemolysis and tumoricidal action. PLoS One. 2013; 8(9):e68390. Published 2013 Sep 11. Http://doi:10.1371/journal.pone.0068390.
2. Patel SG, Karlitz JJ, Yen T, Lieu CH, Boland CR. The rising tide of early-onset colorectal cancer: a comprehensive review of epidemiology, clinical features, biology, risk factors, prevention, and early detection. Lancet Gastroenterol Hepatol. 2022; 7(3), 262-274. https://doi.org/10.1016/S2468-1253(21)00426-X.
3. Rahimi F, Rezayatmand R, Tabesh E, Tohidinik HR, Hemami MR, Ravankhah Z, et al. Incidence of colorectal cancer in Iran: A systematic review and meta-analysis. J Res Med Sci. 2024; 29(1), 65. https://doi.org/10.4103/jrms.jrms11023.
4. Darbandi M, Khorrami Z, Karamoozian A, Aboubakri O, Miryan M, Rezakhani L, et al. A comparison of the burden of cancers between 1990 and 2019 in Iran: A national and subnational study. PLoS One. 2025; 20(2):e0309699. https://doi.org/10.1371/journal.pone.0309699.
5. Jafari A, Hosseini FA, Jalali, FS. A systematic review of the economic burden of colorectal cancer. Health Sci. Rep. 2024; 7(8), e70002. https://doi.org/10.1002/hsr2.70002.
6. Asma ST, Acaroz U, Imre K, Morar A, Shah SRA, Hussain SZ, et al. Natural Products/Bioactive Compounds as a Source of Anticancer Drugs. Cancers. 2022; 14(24):6203. https://doi.org/10.3390/cancers14246203.
7. Håkansson A, Zhivotovsky B, Orrenius S, Sabharwal H, Svanborg C. Apoptosis induced by a human milk protein. Proc Natl Acad Sci U S A. 1995; 92(17):8064-8. https://doi.org/10.1073/pnas.92.17.8064.
8. Wang M, Liu X, Chen T, Cheng X, Xiao H, Meng X, Jiang Y. Inhibition and potential treatment of colorectal cancer by natural compounds via various signaling pathways. Front Oncol. 2022; 12:956793.9. https://doi.org/10.3389/fonc.2022.956793.
9. Islam MR, Akash S, Rahman MM, Nowrin FT, Akter T, Shohag S, et al. Colon cancer and colorectal cancer: Prevention and treatment by potential natural products. Chem Biol Interact. 2022; 368:110170. https://doi.org/10.1016/j.cbi.2022.110170.
10. Zacatecas Z, Campus, II. Perspective of the Use of Hamlet as Palliative in Patients Undergoing Chemotherapy. Acta Sci. Med. Sci. 2024; 8(7). https://doi.org/10.31080/ASMS.2024.08.1842
11. Ho JC, Nadeem A, Svanborg C. HAMLET–A protein-lipid complex with broad tumoricidal activity. Biochem Biophys Res Commun. 2017; 482(3), 454-458. https://doi.org/10.1016/j.bbrc.2016.10.092.
12. Žilinskas J, Stukas D, Jasukaitienė A, Šapauskienė J, Banienė R, Trumbeckaitė S, et al. HAMLET effect on cell death and mitochondrial respiration in colorectal cancer cell lines with KRAS/BRAF mutations. J. Cancer Res. Clin. Oncol. 2023; 149(11):8619-8630. https://doi.org/10.1007/s00432-023-04777-0
13. Žilinskas J. HAMLET Complex and FOLFOX Chemotherapy Impact on Colorectal Cancer Cells and Tissues with Different KRAS/BRAF Mutations. Doctoral dissertation, Lithuanian University of Health Sciences (Lithuania)). 2024.
14. Babazadeh M, Zamani M, Mehrbod P, Mokarram P. The Comparative Role of BAMLET and 5-Fluorouracil in Colorectal Cancer Cells by Targeting WNT/β-Catenin Pathway.Int J Cancer Manag. 2022; 15(9):e123140. https://doi.org/10.5812/ijcm-123140.
15. Zhao H, Ming T, Tang S, Ren S, Yang H, Liu M, et al. Wnt signaling in colorectal cancer: pathogenic role and therapeutic target. Mol Cancer. 2022; 21(1), 144. https://doi.org/10.1186/s12943-022-01616-7.
16. Behrouj H, Mokarram P. BAMLET (Bovine α-lactalbumin made lethal to tumor cells) inhibits autophagy flux and induces apoptosis via down-regulation of protein kinase CK1α and attenuation of the AKT/p-ß-catenin (S552) pathway in RAS-mutated human colorectal HCT 116 cells. Iran J Basic Med Sci. 2023; 26(10):1212-1219. https://doi.org/10.22038/IJBMS.2023.69343.15114.
17. Puthia M, Storm P, Nadeem A, Hsiung S, Svanborg C. Prevention and treatment of colon cancer by peroral administration of HAMLET (human α-lactalbumin made lethal to tumour cells). Gut. 2014; 63(1):131-142. https://doi.org/10.1136/gutjnl-2012-303715.
18. Payandeh Z, Khalili S, Somi MH, Mard-Soltani M, Baghbanzadeh A, Hajiasgharzadeh K, et al. PD-1/PD-L1-dependent immune response in colorectal cancer. J. Cell. Physiol. 2020; 235(7-8), 5461–5475. https://doi.org/10.1002/jcp.29494.
19. Dosset M, Vargas TR, Lagrange A, Boidot R, Végran F, Roussey A, et al. PD-1/PD-L1 pathway: an adaptive immune resistance mechanism to immunogenic chemotherapy in colorectal cancer. Oncoimmunol, 2018; 7(6), e1433981. https://doi.org/10.1080/2162402X.2018.1433981.
20. Tran HT, Wan MLY, Ambite I, Cavalera M, Grossi M, Háček J, et al. BAMLET administration via drinking water inhibits intestinal tumor development and promotes long-term health. Sci Rep. 2024; 14, 3838. https://doi.org/10.1038/s41598-024-54040-w.
21. Jiang H, Pang J, Li T, Akofala A, Zhou X, Yi C, et al. PD-1 regulates the anti-tumor immune function of macrophages through JAK2-STAT3 signaling pathway in colorectal cancer tumor microenvironment. J Transl Med. 2025; 23(1):502. https://doi.org/10.1186/s12967-025-06469-4
22. & Svanborg, C. (2008). Apoptosis and tumor cell death in response to HAMLET (human α-lactalbumin made lethal to tumor cells). Bioactive components of milk, 217-240.
23. Hallgren O, Aits S, Brest P, Gustafsson L, Mossberg AK, Wullt B, et al. Apoptosis and Tumor Cell Death in Response to HAMLET (Human α-Lactalbumin Made Lethal to Tumor Cells). In: Bösze, Z. (eds) Bioactive Components of Milk. Advances in Experimental Medicine and Biology, vol 606. Springer, New York, NY. 2008; https://doi.org/10.1007/978-0-387-74087-4_8.
24. Hallgren O, Gustafsson L, Irjala H, Selivanova G, Orrenius S, Svanborg C. HAMLET triggers apoptosis but tumor cell death is independent of caspases, Bcl-2 and p53. Apoptosis. 2006; 11, 221–233. https://doi.org/10.1007/s10495-006-3607.
25. Žilinskas J, Stukas D, Jasukaitienė A, Žievytė I, Balion Z, Šapauskienė J, et al. Assessing the Therapeutic Impacts of HAMLET and FOLFOX on BRAF-Mutated Colorectal Cancer: A Study of Cancer Cell Survival and Mitochondrial Dynamics in Vitro and Ex Vivo. Medicina. 2024; 60(1):142. https://doi.org/10.3390/medicina60010142.
26. Babazadeh M, Zamani M, Mehrbod P, Mokarram P. Stemness targeting of colorectal cell lines mediated by BAMLET and 5-Flourouracil. Biochem Biophys Res Commun. 2023; 664:136-141. https://doi.org/10.1016/j.bbrc.2023.04.010.
27. Rammer P, Groth-Pedersen L, Kirkegaard T, Daugaard M, Rytter A, Szyniarowski P, et al. BAMLET activates a lysosomal cell death program in cancer cells. Mol. Cancer Ther. 2010; 9(1), 24-32. https://doi.org/10.1158/1535-7163.mct-09-0559.
28. Wu X, Que H, Li Q, Wei X. Wnt/β-catenin mediated signaling pathways in cancer: recent advances, and applications in cancer therapy. Mol. Cancer. 2025; 24(1):171. Published 2025 Jun 10. http://doi:10.1186/s12943-025-02363-1.
29. Yang Z, Manshour N, He X, Epp M, Unruh J, Mao X, et al. Overcoming Immune Escape Via Targeting Akt-Wnt/β-Catenin By Artificial-Intelligence-Powered Peptide-Producing mRNAs. 2024; Blood, 144, 1062. https://doi.org/10.1182/blood-2024-200368.
30. Hoque M, Dave S, Gupta P, Saleemuddin M. Oleic acid may be the key contributor in the BAMLET-induced erythrocyte hemolysis and tumoricidal action. PLoS One. 2013; 8(9):e68390. Published 2013 Sep 11. Http://doi:10.1371/journal.pone.0068390.
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| Issue | Vol 3 No 3 (2025) | |
| Section | Review Article(s) | |
| Keywords | ||
| α-Lactalbumin Oleic Acid Colorectal Cancer Antitumor Activity Naturally Derived Complexes HAMLET BAMLET | ||
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How to Cite
1.
Babazadeh M. Investigating the Antitumor Effects of α-Lactalbumin–Oleic Acid Complexes (HAMLET and BAMLET) in Colorectal Cancer: A Systematic Perspective. ABI. 2025;3(3):125-132.
